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Architecture of an HIV-1 reverse transcriptase initiation complex
Reverse transcription of the HIV-1 RNA genome into double-stranded DNA is a central step in infection(1) and a common target of antiretrovirals(2). The reaction is catalyzed by viral reverse transcriptase (RT)(3,4) that is packaged in an infectious virion along with 2 copies of dimeric viral genomic...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5934294/ https://www.ncbi.nlm.nih.gov/pubmed/29695867 http://dx.doi.org/10.1038/s41586-018-0055-9 |
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author | Larsen, Kevin P. Mathiharan, Yamuna Kalyani Kappel, Kalli Coey, Aaron Chen, Dong-Hua Barrero, Daniel Madigan, Lauren Puglisi, Joseph D. Skiniotis, Georgios Puglisi, Elisabetta Viani |
author_facet | Larsen, Kevin P. Mathiharan, Yamuna Kalyani Kappel, Kalli Coey, Aaron Chen, Dong-Hua Barrero, Daniel Madigan, Lauren Puglisi, Joseph D. Skiniotis, Georgios Puglisi, Elisabetta Viani |
author_sort | Larsen, Kevin P. |
collection | PubMed |
description | Reverse transcription of the HIV-1 RNA genome into double-stranded DNA is a central step in infection(1) and a common target of antiretrovirals(2). The reaction is catalyzed by viral reverse transcriptase (RT)(3,4) that is packaged in an infectious virion along with 2 copies of dimeric viral genomic RNA(5) and host tRNA(Lys)(3), which acts as a primer for initiation of reverse transcription(6,7). Upon viral entry, initiation is slow and non-processive compared to elongation(8,9). Despite extensive efforts, the structural basis for RT function during initiation has remained a mystery. Here we apply cryo-electron microscopy (cryo-EM) to determine a three-dimensional structure of the HIV-1 RT initiation complex. RT is in an inactive polymerase conformation with open fingers and thumb and with the nucleic acid primer-template complex shifted away from the active site. The primer binding site (PBS) helix formed between tRNA(Lys)(3) and HIV-1 RNA lies in the cleft of RT and is extended by additional pairing interactions. The 5′ end of the tRNA refolds and stacks on the PBS to create a long helical structure, while the remaining viral RNA forms two helical stems positioned above the RT active site, with a linker that connects these helices to the RNase H region of the PBS. Our results illustrate how RNA structure in the initiation complex alters RT conformation to decrease activity, highlighting a potential target for drug action. |
format | Online Article Text |
id | pubmed-5934294 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
record_format | MEDLINE/PubMed |
spelling | pubmed-59342942018-10-25 Architecture of an HIV-1 reverse transcriptase initiation complex Larsen, Kevin P. Mathiharan, Yamuna Kalyani Kappel, Kalli Coey, Aaron Chen, Dong-Hua Barrero, Daniel Madigan, Lauren Puglisi, Joseph D. Skiniotis, Georgios Puglisi, Elisabetta Viani Nature Article Reverse transcription of the HIV-1 RNA genome into double-stranded DNA is a central step in infection(1) and a common target of antiretrovirals(2). The reaction is catalyzed by viral reverse transcriptase (RT)(3,4) that is packaged in an infectious virion along with 2 copies of dimeric viral genomic RNA(5) and host tRNA(Lys)(3), which acts as a primer for initiation of reverse transcription(6,7). Upon viral entry, initiation is slow and non-processive compared to elongation(8,9). Despite extensive efforts, the structural basis for RT function during initiation has remained a mystery. Here we apply cryo-electron microscopy (cryo-EM) to determine a three-dimensional structure of the HIV-1 RT initiation complex. RT is in an inactive polymerase conformation with open fingers and thumb and with the nucleic acid primer-template complex shifted away from the active site. The primer binding site (PBS) helix formed between tRNA(Lys)(3) and HIV-1 RNA lies in the cleft of RT and is extended by additional pairing interactions. The 5′ end of the tRNA refolds and stacks on the PBS to create a long helical structure, while the remaining viral RNA forms two helical stems positioned above the RT active site, with a linker that connects these helices to the RNase H region of the PBS. Our results illustrate how RNA structure in the initiation complex alters RT conformation to decrease activity, highlighting a potential target for drug action. 2018-04-25 2018-05 /pmc/articles/PMC5934294/ /pubmed/29695867 http://dx.doi.org/10.1038/s41586-018-0055-9 Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms Reprints and permissions information is available at www.nature.com/reprints. |
spellingShingle | Article Larsen, Kevin P. Mathiharan, Yamuna Kalyani Kappel, Kalli Coey, Aaron Chen, Dong-Hua Barrero, Daniel Madigan, Lauren Puglisi, Joseph D. Skiniotis, Georgios Puglisi, Elisabetta Viani Architecture of an HIV-1 reverse transcriptase initiation complex |
title | Architecture of an HIV-1 reverse transcriptase initiation complex |
title_full | Architecture of an HIV-1 reverse transcriptase initiation complex |
title_fullStr | Architecture of an HIV-1 reverse transcriptase initiation complex |
title_full_unstemmed | Architecture of an HIV-1 reverse transcriptase initiation complex |
title_short | Architecture of an HIV-1 reverse transcriptase initiation complex |
title_sort | architecture of an hiv-1 reverse transcriptase initiation complex |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5934294/ https://www.ncbi.nlm.nih.gov/pubmed/29695867 http://dx.doi.org/10.1038/s41586-018-0055-9 |
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